Welcome to Scribd, the world's digital library. Read, publish, and share books and documents. See more
Download
Standard view
Full view
of .
Look up keyword
Like this
1Activity
0 of .
Results for:
No results containing your search query
P. 1
Asawalam et al

Asawalam et al

Ratings: (0)|Views: 40|Likes:
Published by tissueculture

More info:

Published by: tissueculture on Mar 04, 2010
Copyright:Attribution Non-commercial

Availability:

Read on Scribd mobile: iPhone, iPad and Android.
download as PDF, TXT or read online from Scribd
See more
See less

03/04/2010

pdf

text

original

 
 
African Journal of Biotechnology Vol. 7 (20), pp. 3771-3776, 20 October, 2008Available online at http://www.academicjournals.org/AJBISSN 1684–5315 © 2008 Academic Journals
 
Full Length Research Paper 
Essential oil of
Ocimum grattissimum 
(Labiatae) as
Sitophilus zeamais 
(Coleoptera: Curculionidae)
 
protectant
E. F. Asawalam
1,2
*, S. O. Emosairue
2
and A. Hassanali
1
 
1
Behavioural and Chemical Ecology Department, International Centre of Insect Physiology and Ecology P. O. Box 30772-00100 Nairobi, Kenya.
2
Department of Plant Health Management, Michael Okpara University of Agriculture, Umudike P. M. B. 7267 Umuahia,Abia state Nigeria.
Accepted 27 August, 2008
Ocimum grattissimum 
L. (Labiatae) leaves are widely eaten as a vegetable in Nigeria, and in the easternparts, are traditionally used in post-harvest protection and relieving stomach aches. The effect of theessential oil of
O. grattissimum 
leaves on
Sitophilus zeamais 
(Motschulsky) (Coleoptera: Curculionidae)was assessed for repellency, mortality, progeny emergence and maize damage in the laboratory. The oilwas found to be moderately repellent to the maize weevil and induced high mortality in the weevils. Inaddition, grains treated with the essential oil showed significant reduction in the number of progenyderived from surviving
S. zeamais 
. There was no observable feeding damage on grains treated with thehighest dosage of the essential oil extract. Gas chromatography-linked mass spectrometry (GC-MS)and GC co-injections with authentic samples showed the presence of the following major constituents:thymol (32.7%), paracymene (25.4%),
-terpinene (10.8%),
 
-selinene (4.5%), phellandrene (3.9%) and
 
-myrcene (3.1%). The results provide a scientific rationale for the use of the plant in post-harvestprotection.Key words:
 
Ocimum grattissimum 
, essential oil,
Sitophilus zeamais,
maize, repellency.
INTRODUCTION
Maize (
Zea mays 
L.) is one of the foremost cerealscultivated in the world today (Purseglove, 1975;Rouanet, 1992) and it is a major source of dietarycarbohydrate in the tropics (Wudiri and Fatobi, 1992).The maize weevil,
Sitophilus zeamais 
(Motschulsky)(Coleoptera:Curculionidae), is a serious pest of maizeand it is capable of developing on all other cereal grainsand cereal products (Walgenbach and Burkholder, 1986;Tipping et al., 1987). Initial infestation of maize grainoccurs in the field just before harvest and insects arecarried into the store where the population builds uprapidly (Appert, 1987; Adedire and Lajide, 2003). The
*Corresponding author. E-mail: elechiasw@yahoo.com,easawalam@icipe.org. Tel: +2348039695157.
 huge post-harvest losses and quality deteriorationcaused by this pest is a major obstacle to achieving foodsecurity in developing countries (Rouanet, 1992).The efficient and effective control of storage insectslike
S. zeamais 
has centered mainly on the use ofsynthetic insecticides (Menn, 1983; Redlinger et al.,1988). However, many problems are associated withthese chemicals, such as the development of insectresistance, toxic residues in food, workers’ safety, andhigh cost of procurement (Sighamony et al., 1990).These problems have necessitated search for alterna-tive eco-friendly insect pest control methods amongstwhich are the use of botanical pesticides (Cobbinah andAppiah-Kwarteng, 1989; Hassanali et al., 1990; Niber,1994; Jembere et al., 1995; Bekele et al., 1996; Lajide etal, 1998; Asawalam and Adesiyan, 2001; Asawalam andAdesiyan, 2002; Bekele, 2002; Asawalam and Arukwe,
 
 
3772 Afr. J. Biotechnol.2004).
Ocimum grattissimum 
L
.
(African Curry plant) is a lowgrowing shrubby species in the family Labiatae. All thespecies in the genus contain strongly scented essentialoils. It is grown as pot herbs for local medicines andexists in diversity of forms and cultivars (Schippers,2000). The powders and essential oils of
Ocimum 
 species have been widely used for control of insect pestsespecially storage insect pests (Oparaeke et al
.
, 2002). Itis a slightly hairy annual with much branched angularstems carrying opposite, ovate leaves which are usuallyless than 1 cm long and borne on fairly long petioles. Ineastern Nigeria, it serves as a source of stimulant andcondiment in soup. Medicinally, it is used for thetreatment of stomach ache, sores and in management ofbabies cord after delivery among the Igbos of Nigeria(Ijeh et al
.
, 2004).The aim of the present study was to document thebiological effects of the essential oil on the maize weeviland to analyse its composition in order to assess itsrelative safety.
MATERIALS AND METHODS
Sitophilus zeamais 
culture
S. zeamais 
was cultured in the laboratory at 27 ± 2ºC, 60 - 65%R.H and 12 h : 12 h light : dark regime. The food media used wasinsecticide-free whole maize grains purchased from a local market(Dikomba) outside Nairobi in Kenya. Fifty pairs of
S. zeamais 
wereplaced in 1 L glass jar containing 400 g of maize grains. The jarswere then covered with nylon mesh held in place with rubberbands. Grains were disinfested in the oven at 40
o
C for 4 h(Jembere et al
.
, 1995) and kept in the laboratory before use.
Plant collection and isolation of their essential oil
O. grattissimum 
 
leaves were collected from Umudike, Nigeria inJanuary 2006. The identity of the plant was confirmed at theherbarium of Michael Okpara University of Agriculture, Umudike,Nigeria, before using them for the present study. Plants were air-dried in a well-ventilated area for five days before extraction.Voucher specimens are kept at the University herbarium.
 
The essential oil was extracted by steam distillation usingClavenger apparatus (Guenter, 1949). The condensing oils werecollected in n-hexane (Aldrich HPLC grade) and the solution wasfiltered and exposed to anhydrous sodium sulphate to remove anytraces of water. Hexane was then removed by distillation at 60
o
Cfrom 'Contes' Short Path distillation apparatus and the oil collectedand weighed, and stored in small amber-coloured vials.
Analysis of essential oil
Gas chromatographic (GC) analyses were performed on a capillarygas-chromatograph Hewlett Packard (HP) 5890 Series II equippedwith a split-less capillary injector system, 50 m x 0.20 mm (i.d.)cross-linked with HP-ultra 1methylsilicone 0.33 µm (film thickness)capillary column, and Flame Ionization Detector (FID) coupled toHP 3393A Series II integrator. The integrator was used to calculatethe peak areas. The carrier gas was nitrogen at a flow rate of 0.84ml/min. The temperature programme comprised of an initialtemperature of 40
o
C (0 min) to 90
o
C at 7
o
C/min, a hold at thistemperature for 5 min, then to 115 at 3
o
C/min followed by anotherhold for 5 min, and finally to 280
o
C at 4
o
C/min where it wasmaintained for 20 min.Gas chromatography–linked mass Spectrometry (GC-MS)analysis was carried out on a HP 8060 Series II gas chromatographcoupled to VG Platform II Mass Spectrometer (manufactured byMicromass, UK, formerly known as VG Biotech). The MS wasoperated in the Electron impact mode (EI) at 70 eV and anemission current of 200
µ
A. The temperature of source was held at180
o
C and the multiplier voltage at 300 V. The pressure of the ionsource and MS detector were held at 9.4 x 10
-6
and 9.4 x 10
-6
mbar,respectively. The MS had a scan cycle of 1.5 s (scan duration of 1 sand inter-scan delay, 0.5 s). The mass and scan range was set atm/z 1 - 1400 and 38 - 650, respectively. The instrument wascalibrated using heptacosafluorotributyl amine, [CF
3
(CF
2
)
3
]
3
N,(Apollo Scientific Ltd., UK). Column (film thickness 0.5
µ
m)temperature was programmed as in the case of GC analysis. AllGC-MS analyses were made in the splitless mode with helium asthe carrier gas. Preliminary identification of constituents was basedon computer matching components of mass spectral data againstthe standard Wiley and NIST library spectra, constituted fromspectra of pure substances and components of the known essentialoils, and literature MS data. They were confirmed by their GCretention time comparison with those of reference compounds,peak enhancement as well as co-injection /co-elution with authenticsamples. The samples used were obtained from Aldrich ChemicalsUK. Relative proportion of the essential oil was computed in eachcase from GC-MS peak areas.
Repellency
Repellent action of the essential oil against
S. zeamais 
wasassessed in a choice bioassay system at 27 ± 2
o
C and 60 - 65%R.H. previously reported by Bekele et al. (1996). It consisted oftwo 1 L glass jars connected together at their rims by means of a30 x 10 cm nylon mesh tube. A 5.0 cm diameter circular hole wascut at the middle of the mesh for the introduction of test insects.250 g of maize were put into each glass jars. Grains in one jarwere treated with the essential oil at the rate of 0.012, 0.06 and0.3% (30, 150 and 750 mg/250 g) while untreated grains in theother jar acted as control. Twenty-five adults of
S. zeamais 
wer
introduced into the nylon mesh tube through the circular hole bymeans of a 5 cm diameter funnel. The number of insects presentat the control (N
C
) and treated (N
T
) jars were recorded after 1-hour exposure. All repellency assays were replicated four times.Percent repellency (PR) values were computed from the formula:PR = [N
C
-N
T
/ N
C
+N
T
] × 100.PR data were analyzed using ANOVA after arcsinetransformation.
Effect of essential oil on mortality
Essential oil was applied to the grains at the rate of 0.012, 0.06and 0.3% dissolved in 10 ml of 95% n-hexane and shakenthoroughly to ensure uniform distribution over grain surface.Treated grains were kept for 24 h to allow the hexane to evapo-rate completely before bioassays were conducted. Two blankcontrols were run concurrently consisting of hexane treated grainand untreated grain. Ten pairs of 5/7-day-old
S. zeamais 
adults
 
 
Asawalam et al. 3773
Table 1.
Biological activity of different doses
 
of
O. grattissimum 
essential oil against
S. zeamais 
.
Treatments Mean repellency (%) Mean no of F1 progeny Mean weight loss (%)
30 mg (0.012%) 36.5 ± 1.6
c
119.25 ± 2.6
c
14.8 ± 1.6
c
 150 mg (0.06%) 58.75 ± 2.1
b
82.75 ± 3.5
b
8.35 ± 2.1
b
 750 mg 0.6%) 77.5 ± 1.7
a
0 ± 0
a
0 ± 0
a
 Hexane 0 ± 0
d
152.75 ± 3.2
d
22.1 ± 1.9
d
 Untreated 0 ± 0
d
156.28 ± 2.6
d
23.15 ± 1.8
d
 
Values are mean ± SE (Standard error).Treatment means (average of four replicates) within each column
 
followed by the same letter are not significantlydifferent
 
from each other
 
at
 
5% level of probability according to Student –Newman – Keuls (SNK) test.
R
2
= 0.94R
2
= 0.95R
2
= 0.86
010203040506070809010002468
Days after treatment
   C  u  m  u   l  a   t   i  v  e   %   m  o  r   t  a   l   i   t  y
30mg150mg750mgy = aLn(x) + b
 
Figure 1.
Cumulative mean (%) mortality of
S. zeamais 
in maizegrains treated with different concentrations of
O. grattissimum 
 essential oil.were introduced into each of the treated and untreated jars. The jars were covered with nylon mesh held with rubber bands. Eachtreatment was replicated four times. The experiment was arrangedin completely randomized design in the laboratory.Number of dead insects in each vial was counted daily for 7days to estimate maize weevil mortality. The percentage mortalitywas corrected using Abbott’s (1925) formulaPT
 
= (P
o
– P
c
) / (100 - P
c
)Where P
T
= corrected mortality (%), P
o
= observed mortality (%), P
c
= control mortality (%).
Effect of essential oil on progeny production
10 pairs of
S. zeamais 
adults were introduced into each treated (atthe same doses as above) and untreated grains. Insectssubsequently emerging were counted to estimate F
I
progenyproduction. Counting was stopped after 33 days to avoidoverlapping of generation.
Effect of essential oil on maize damage by the weevil
Damage assessment was carried out on treated (at the 3 doses asabove) and untreated grains. Samples of 100 grains were takenfrom each jar and the number of undamaged and damaged (grainswith characteristic holes) grains were counted and weighed.Percentage weight loss was calculated using the formula:Weight loss (%) = [UaNv - (U+D)] / UaN × 100Where U = weight of undamaged fraction in the sample, N = totalnumber of grains in the sample, Ua = average weight of oneundamaged grain, D = weight of damaged fraction in the sample(FAO, 1985).
Statistical analysis
Data obtained were subjected to analysis of variance (ANOVA)using a general linear model procedure (SAS, 2002) and meanswere separated by Student Newman-Keuls (SNK) test at P < 0.05.
RESULTSRepellency effect of the essential oil on
S. zeamais 
The mean repellency values of the essential oil at thethree dose levels against
S. zeamais 
are provided inTable 1. All the dosages were repellent to
S. zeamais 
ina dose-response manner. Analysis of variance indicatedsignificant differences (P < 0.05) between weevilresponses to the three dosages tested.
Adult mortality in grains
Figure 1 also shows the cumulative mean percentagemortality of
S. zeamais 
in maize grains treated withdifferent concentrations of
O. grattissimum 
 
essential oil.All treatments with essential oil showed significant levelof mortality with the highest dose (corresponding to0.3%) inducing 82% mortality after 7 days treatment.There was no significant mortality in the untreatedcontrols.
Effect of essential oil on progeny production
The number of progeny produced by
S. zeamais 
in un-

You're Reading a Free Preview

Download
scribd
/*********** DO NOT ALTER ANYTHING BELOW THIS LINE ! ************/ var s_code=s.t();if(s_code)document.write(s_code)//-->